PROJECT SUMMARY Gonorrhea is caused by the bacterium Neisseria gonorrhoeae and is the second most reported sexually transmitted infection (STI) in the world. If left untreated, complications may include pelvic inflammatory disease, newborn blindness, infertility and increased risk of infection by other STDs, including HIV. The Centers for Disease Control and Prevention (CDC) estimates that ~820,000 new gonococcal infections occur in the U.S. each year with 570,000 among young people 15-24 years old. Medications to treat gonorrhea have been around for decades but the organism has grown resistant to virtually every medication used to treat it. The CDC recommends treatment changes when surveillance data indicate a ?5% resistance to the class of antimicrobial being used at the time. Currently, dual treatment with ceftriaxone (cephalosporin) with azithromycin (macrolide) are recommended. Since treatment recommendations are based on ?5% resistance, most circulating isolates appear to still be sensitive to previously-recommended antimicrobials. Re-introducing these antimicrobials into clinical use may allow for the continued successful treatment of gonorrhea even if cephalosporin resistance becomes widespread. Culture-based agar dilution tests are the gold standard for determining the susceptibility of N. gonorrhoeae to antibiotics. However, it is unacceptably slow (~2 days) and laborious. Faster genotypic approaches are available that involve the detection of gene mutations known to confer resistance (e.g., NGSNP). However, these approaches are limiting, as they require prior knowledge of resistance mechanisms. To select the most effective therapy, antimicrobial susceptibility diagnostics are needed to characterize phenotypes of infecting N. gonorrhoeae strains. These tests can guide treatment at the point of care (POC), and facilitate antibiotic stewardship. Lynntech is proposing to develop a test based on the molecular responses of N. gonorrhoeae to antibiotic exposure. Molecular responses upon exposure to antibiotics can be detected long before phenotypic changes in growth can be observed. Measuring molecular changes in N. gonorrhoeae to antibiotic exposure will provide a rapid test that can be performed at the POC, such as a clinic. Lynntech?s assay will be rapid (~ 1 hour) and will be able to identify N. gonorrhoeae as well as determine the susceptibility of the infecting strain to at least three classes of antibiotics (quinolones, macrolides, cephalosporins) directly from the patient sample (e.g., urine, urethral/vaginal/cervical swab). Our assay will be self-contained, require minimal hands-on time and will provide easy to interpret results for the clinician.